The European eel (Anguilla anguilla) is a commercially valued species for aquaculture. Over the past decades, it has experienced a drastic reduction in its natural stocks. Thus, breeding in captivity is considered essential, nowadays, to guarantee the eel aquaculture and to reduce pressure on natural populations. Traditionally, the European eel has been sexually matured by means of weekly hormonal injections, which cause stress to the fish. The purpose of this research study was to assess the use of osmotic pumps as a new method to induce sexual maturation in male and female European eels, without the weekly injection. The control groups were treated with weekly hormone injections (recombinant human chorionic gonadotropin for males and carp pituitary extract for females), and the implanted groups were treated with osmotic pumps (ALZET^® osmotic pumps) loaded with the respective hormones. Regarding male European eels, this study shows that the use of controlled release systems was able to induce the maturation and spermiation, but without the necessary capacity to produce enough gametes with acceptable quality parameters that could meet the needs of a commercial eel hatchery. Concerning female European eels, the study demonstrates that the use of osmotic pumps loaded with CPE became an effective method, generating early maturations (4 to 10 weeks) in 50% of the females, so this method could become a viable alternative for eel hatchery procedures.

Identification of specific molecular markers for spermatogonial stem cells in teleost is crucial for enhancing the efficacy of reproductive biotechnologies in aquaculture, such as transplantation and surrogate production in fishes. Since it is not yet possible to distinguish spermatogonial stem cells of European eel (Anguilla anguilla) using specific molecular markers, we isolated spermatogonial cells from immature European eels to find these potential markers. We attempted this by studying three candidate genes: vasa, nanos2, and dnd1. Two vasa (vasa1 and vasa2) genes, nanos2, and dnd1 were identified, characterized, and studied in the muscle, testis, and isolated spermatogonia. Our results showed that vasa1 and vasa2 had the highest levels of expression when measured by qPCR. In situ hybridization and immunochemistry assays showed that the four genes were localized explicitly in type A spermatogonia. However, vasa1 and vasa2 exhibited stronger signals in the immature testicular tissue than the other two potential markers. According to this, vasa1 and vasa2 were found to be the most effective markers for spermatogonial cells in the European eel.

The European eel encounters challenges in achieving sexual maturation in captivity, which has been a concern for researchers. This study explores surrogate broodstock technology as an alternative approach for eel production. The present study aimed to evaluate zebrafish and European sea bass as potential recipients for European eel spermatogonia transplantation, given the abundance of eel type A spermatogonia (SPGA). Immature European eel testes were dissected and maintained at 4 °C or cryopreserved. SPGA were obtained by dissociation of fresh or post-thawed tissue, employing an enzymatic solution, and then labelled with fluorescent membrane marker PKH26. SPGA from fresh tissue were transplanted into wild-type zebrafish larvae and triploid European sea bass larvae, while SPGA from cryopreserved testis were transplanted into vasa::egfp transgenic zebrafish larvae. One-and-a-half months post-transplantation (mpt), fluorescent donor cells were not detected in the gonads of zebrafish or European sea bass. Molecular qPCR analyses at 1.5 or 6 mpt did not reveal European eel-specific gene expression in the gonads of any transplanted fish. The findings suggest that the gonadal microenvironments of zebrafish and European sea bass are unsuitable for the development of European eel spermatogonia, highlighting distinctive spermatogonial stem cell migration mechanisms within teleost species